Journal: mAbs

Article Title: Improved antibody pharmacokinetics by disruption of contiguous positive surface potential and charge reduction using alternate human framework.

doi: 10.1080/19420862.2023.2232087

Figure Lengend Snippet: Figure 2. Investigation of factors contributing to fast clearance of ACI-5891.1. (a) Human FcRn (hFcRn) binding of ACI-5891.1 at pH 6.0 (left panel) and pH 7.4 (right panel). SPR measurement performed using antibody concentrations of 3000 nM, 1000 nM, 333.3 nM, 111.0 nM, and 37.0 nM as depicted. Sensorgrams are shown in black lines. (b) Non-specific binding of ACI-5891.1 and ACI-5891.5 by heparin and insulin ELISA. Mean signals for both targets are reported as nonspecific binding score. (c) Surface representation of ACI-5891.1 Fv model colored according to Eisenberg’s scale of hydrophobicity (shown at the bottom of panel c). Intense red indicates the most hydrophobic residue while white represents the most hydrophilic residue.

Article Snippet: Biotinylated hFcRn (AcroBiosystems, cat # FCMH82W4) was immobilized on flow-cell 2 (fc2) of flowchannels 1 to 8 of a streptavidin (SA) sensor chip (Cytiva, cat# BR-1005-31) to reach a final surface density of approximatively 150 RU.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Residue

Journal: Communications Biology

Article Title: Interaction of clinical-stage antibodies with heme predicts their physiochemical and binding qualities

doi: 10.1038/s42003-021-01931-7

Figure Lengend Snippet: a Real-time binding profiles of selected heme-binding and heme-sensitive Abs to surface-immobilized human recombinant FcRn. The interaction analyses were performed with native and Abs exposed to an excess of hemin. The black lines depict the binding profiles obtained after injection of serial dilutions of monoclonal Abs, native, and after heme exposure (25–0.195 nM). The red lines depict the fits of data obtained by global analysis using the Langmuir kinetic model. All interaction analyses were performed at 25 °C. b Values of the kinetic rate constants of association (top panel), dissociation (middle panel), and equilibrium dissociation constant (bottom panel) for binding of native and heme-exposed monoclonal Abs to FcRn. The kinetics data were obtained after analyses of the real-time interaction profiles shown in ( a ).

Article Snippet: Biotinylated recombinant human FcRn (Amsbio LLC, Cambridge, MA) was diluted to 5 μg/ml in running buffer—100 mM Tris pH 6 (adjusted by addition of dry citric acid), 100 mM NaCl, 5 % Glycerol, and 0.1% Tween 20—and injected with a flow rate of 10 μl/min over the pre-activated sensor chip surface, coated with streptavidin by the manufacturer (SA chip, Biacore).

Techniques: Binding Assay, Recombinant, Injection